1. PI3K/Akt/mTOR
    Stem Cell/Wnt
  2. GSK-3

CHIR-99021 trihydrochloride (Synonyms: CT99021 trihydrochloride)

Cat. No.: HY-10182B Purity: 99.02%
Handling Instructions

CHIR-99021 trihydrochloride is a GSK-3α/β inhibitor with IC50 of 10 nM/6.7 nM; > 500-fold selectivity for GSK-3 versus its closest homologs CDC2 and ERK2, as well as other protein kinases.

For research use only. We do not sell to patients.
CHIR-99021 trihydrochloride Chemical Structure

CHIR-99021 trihydrochloride Chemical Structure

CAS No. : 1782235-14-6

Size Price Stock Quantity
Free Sample (0.5-1 mg)   Apply now  
10 mM * 1 mL in DMSO USD 121 In-stock
5 mg USD 96 In-stock
10 mg USD 156 In-stock
50 mg USD 588 In-stock
100 mg   Get quote  
200 mg   Get quote  

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Customer Review

Other Forms of CHIR-99021 trihydrochloride:

    CHIR-99021 trihydrochloride purchased from MCE. Usage Cited in: Toxicology. 2016 May 20;355-356:31-38.

    Western analysis of β-catenin protein level in zebrafish embryos exposed to EOM in the presence or absence of CH/CHIR (n = 3). Different letters indicate significant differences. Moreover, both CH and CHIR attenuate the EOM-induced changes in the protein level of b-catenin and in EROD activity.

    CHIR-99021 trihydrochloride purchased from MCE. Usage Cited in: Cell Signal. 2016 Mar;28(3):148-56.

    Inhibition of PI3K p110δ signalling, either by PI3KCD-/- or using PI3K p110δ inhibitor PI-3065, significantly blocks LPS-induced GSK3α phosphorylation.

    CHIR-99021 trihydrochloride purchased from MCE. Usage Cited in: Nat Med. 2016 May;22(5):547-56.

    Immunofluorescent staining for α-actinin (ACTN2) and cardiac troponin T (TNNT2) to demonstrate sarcomeric organization in hiPSC–CMs derived from patients who do not experience Doxorubicin–induced cardiotoxicity (DOX) versus those who do experience Doxorubicin-induced cardiotoxicity (DOXTOX) after 24 h treatment with Doxorubicin. Scale bar, 20 µm.
    • Biological Activity

    • Protocol

    • Technical Information

    • Purity & Documentation

    • References


    CHIR-99021 trihydrochloride is a GSK-3α/β inhibitor with IC50 of 10 nM/6.7 nM; > 500-fold selectivity for GSK-3 versus its closest homologs CDC2 and ERK2, as well as other protein kinases.

    IC50 & Target

    IC50: 10 nM/6.7 nM (GSK-3α/β)[1]

    In Vitro

    CHIR 99021inhibits human GSK-3β with Ki values of 9.8 nM[1]. CHIR 99021 is a small organic molecule that inhibits GSK3α and GSK3β by competing for their ATP-binding sites.In vitro kinase assays reveal that CHIR 99021 specifically inhibits GSK3β (IC50=~5 nM) and GSK3α (IC50=~10 nM), with little effect on other kinases[2]. In the presence of CHIR-99021 the viability of the ES-D3 cells is reduced by 24.7% at 2.5 μM, 56.3% at 5 μM, 61.9% at 7.5 μM and 69.2% at 10 μM CHIR-99021 with an IC50 of 4.9 μM[3].

    In Vivo

    In ZDF rats, a single oral dose of CHIR 99021 (16 mg/kg or 48 mg/kg) rapidly lowers plasma glucose, with a maximal reduction of nearly 150 mg/dl 3-4 h after administration[1]. CHIR99021 (2 mg/kg) given once, 4 h before irradiation, significantly improves survival after 14.5 Gy abdominal irradiation (ABI). CHIR99021 treatment significantly blocks crypt apoptosis and accumulation of p-H2AX+ cells, and improves crypt regeneration and villus height. CHIR99021 treatment increases Lgr5+ cell survival by blocking apoptosis, and effectively prevents the reduction of Olfm4, Lgr5 and CD44 as early as 4 h[4].

    Preparing Stock Solutions
    Concentration Volume Mass 1 mg 5 mg 10 mg
    1 mM 1.7400 mL 8.6999 mL 17.3998 mL
    5 mM 0.3480 mL 1.7400 mL 3.4800 mL
    10 mM 0.1740 mL 0.8700 mL 1.7400 mL
    Please refer to the solubility information to select the appropriate solvent.
    Kinase Assay

    Kinases are purified from SF9 cells through use of their His or Glu tag. Glu-tagged proteins are purified, and His-tagged proteins are purified. Kinase assays are performed in 96-well plates with appropriate peptide substrates in a 300-μL reaction buffer (variations on 50 mM Tris-HCl, pH 7.5, 10 mM MgCl2, 1 mM EGTA, 1 mMdithiothreitol, 25 mMβ-glycerophosphate, 1 mM NaF, and 0.01% bovine serum albumin). Peptides has Km values from 1 to 100 μM. CHIR 99021 or CHIR GSKIA is added in 3.5 μL of Me2SO, followed by ATP to a final concentration of 1 μM. After incubation, triplicate 100-μL aliquots are transferred to Combiplate 8 plates containing 100 μL/well of 50 μM ATP and 20 mM EDTA. After 1 hour, the wells are rinsed five times with phosphate-buffered saline, filled with 200 μL of scintillation fluid, sealed, and counted in a scintillation counter 30 min later. All of the steps are at room temperature. The percentage of inhibition is calculated as 100×(inhibitor-no enzyme control)/(Me2SO control-no enzyme control)[2]. MCE has not independently confirmed the accuracy of these methods. They are for reference only.

    Cell Assay

    CHIR 99021 is dissolved in DMSO and stored, and then diluted with appropriate media before use[3].

    The viability of the mouse ES cells is determined after exposure to different concentrations of GSK3 inhibitors for three days using the MTT assay. The decrease of MTT activity is a reliable metabolism-based test for quantifying cell viability; this decrease correlates with the loss of cell viability. 2,000 cells are seeded overnight on gelatine-coated 96-well plates in LIF-containing ES cell medium. On the next day the medium is changed to medium devoid of LIF and with reduced serum and supplemented with 0.1-1 μM BIO, or 1-10 μM SB-216763, CHIR-99021 or CHIR-98014. Basal medium without GSK3 inhibitors or DMSO is used as control. All tested conditions are analyzed in triplicates[3]. MCE has not independently confirmed the accuracy of these methods. They are for reference only.

    Animal Administration

    CHIR 99021 is formulated as solutions in 20 mM citrate-buffered 15% Captisol or as fine suspensions in 0.5% carboxymethylcellulose (Rats)[1].
    CHIR 99021 is prepared in DMSO (Mice)[4].

    Primary hepatocytes from male Sprague Dawley rats that weighed <140 g are prepared and used 1-3 h after isolation. Aliquots of 1×106cells in 1 mL of DMEM/F12 medium plus 0.2% BSA and CHIR 99021(orally at 16 or 48 mg/kg) or controls are incubated in 12-well plates on a low-speed shaker for 30 min at 37°C in a CO2-enriched atmosphere, collected by centrifugation and lysed by freeze/thaw in buffer A plus 0.01% NP40; the GS assay is again performed.
    Mice 6-10 weeks old are used. The PUMA+/+ and PUMA-/- littermates on C57BL/6 background (F10) and Lgr5-EGFP (Lgr5-EGFP-IRES-creERT2) mice are subjected to whole body irradiation (TBI), or abdominal irradiation (ABI). Mice are injected intraperitoneally (i.p.) with 2 mg/kg of CHIR99021 4 h before radiation or 1 mg/kg of SB415286 28 h and 4 h before radiation. Mice are sacrificed to collect small intestines for histology analysis and western blotting. All mice are injected i.p. with 100 mg/kg of BrdU before sacrifice. MCE has not independently confirmed the accuracy of these methods. They are for reference only.

    Molecular Weight




    CAS No.




    Powder -20°C 3 years
      4°C 2 years
    In solvent -80°C 6 months
      -20°C 1 month

    Room temperature in continental US; may vary elsewhere

    Solvent & Solubility

    DMSO: ≥ 32 mg/mL

    * "<1 mg/mL" means slightly soluble or insoluble. "≥" means soluble, but saturation unknown.

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    CHIR-99021 trihydrochloride
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